冰冻切片技术在铁皮石斛显微结构上的应用

doi:10.7525/j.issn.1673-5102.2019.04.016

植物研究 ›› 2019, Vol. 39 ›› Issue (4): 613-618.doi: 10.7525/j.issn.1673-5102.2019.04.016

冰冻切片技术在铁皮石斛显微结构上的应用

蒋素华¹, 梁芳¹, 张燕¹, 周一冉², 郭梦园¹, 袁秀云¹, 崔波¹

1. 郑州师范学院生物工程研究所, 郑州 450044;
2. 郑州大学生命科学学院, 郑州 450002

收稿日期:2019-01-06 出版日期:2019-07-05 发布日期:2019-07-03
通讯作者: 崔波 E-mail:laocuibo@163.com
作者简介:蒋素华(1983-),女,副教授,硕士,主要从事花卉分子生物学研究。
基金资助:
河南省科技厅重大专项（701519，702056）；河南省科技厅科技发展计划项目（172107000003，172102110227）；郑州师范学院科技创新团队支持计划资助（兰花种质资源开发与产业化应用科研创新团队201808）

Cryo-sectioning Technique for Microstructure Studies on Dendrobium officinale

JIANG Su-Hua¹, LIANG Fang¹, ZHANG Yan¹, ZHOU Yi-Ran², GUO Meng-Yuan¹, YUAN Xiu-Yun¹, CUI Bo¹

1. Institute of Bioengineering, Zhengzhou Normal University, Zhengzhou 450044;
2. College of Life Sciences, Zhengzhou University, Zhengzhou 450002

Received:2019-01-06 Online:2019-07-05 Published:2019-07-03
Supported by:
Supported by major projects of Henan province department(701519,702056);Supported by project of scientific development of Henan province department technology and innovation team(172107000003,172102110227);Support Scheme Funding in Zhengzhou Normal University(Orchid germplasm resources development and industrialization applied scientific research innovation team(201808)

摘要/Abstract

摘要： 为了快速高效的观察兰科植物铁皮石斛的显微结构，利用光镜和改进的蔗糖保护——液氮速冻冰冻切片法，观察铁皮石斛根、茎、叶的显微结构。该技术方法是将铁皮石斛器官经过蔗糖磷酸缓冲液保护液处理后抽真空，再经过液氮速冻、包埋、切片、展片观察、染色以及拍照等步骤，制作出铁皮石斛根、茎和叶较完整的显微结构切片。研究结果表明，适合铁皮石斛根的最适条件为：蔗糖质量体积分数为8%、冷凝温度-25℃、切片厚度20 μm；茎的最适条件为：蔗糖质量体积分数为16%、冷凝温度-20℃、切片厚度15 μm；叶的最适条件为：蔗糖质量体积分数为4%、冷凝温度-20℃、切片厚度10 μm。该研究在兰科植物显微结构观察和组织化学研究中将具有广阔的应用前景。

关键词: 铁皮石斛, 蔗糖保护—液氮速冻法, 冰冻切片技术, 显微技术

Abstract: In order to observe the microscopic structure of Dendrobium officinale quickly and efficiently. This study used the light microscope and the improved sucrose protection-liquid nitrogen quick-freezing freezing section method for orchid tissues and organs, Which improved sucrose protection-liquid nitrogen frozen method, observed root, stem and leaf of microscopic structure on D. officinale. The technology and methods treated D. officinale with sucrose phosphate buffer protection fluid,and then Vacuum processing. By liquid nitrogen frozen, embedding, sectioning, exhibition slice observation, staining and photo produced D. officinale roots, stems and leaves a complete microscopic section. The results show that the optimal condition for root was 8% sucrose mass volume fraction, condensation temperature -25℃, slice thickness 20 μm; the optimal condition for stem was 16% sucrose mass volume fraction, frozen temperature -20℃ and slice thickness 15 μm; the optimal condition for leaf was 4% sucrose mass volume fraction, frozen temperature -20℃ and slice thickness 10 μm. The study on the Orchidaceae microstructure observation and histochemical study will have broad prospect of application.

Key words: Dendrobium officinale, frozen section, sucrose-protecting and liquid nitrogen quick frozen method, microstructure

中图分类号:

Q949.71+8.43

蒋素华, 梁芳, 张燕, 周一冉, 郭梦园, 袁秀云, 崔波. 冰冻切片技术在铁皮石斛显微结构上的应用[J]. 植物研究, 2019, 39(4): 613-618.

JIANG Su-Hua, LIANG Fang, ZHANG Yan, ZHOU Yi-Ran, GUO Meng-Yuan, YUAN Xiu-Yun, CUI Bo. Cryo-sectioning Technique for Microstructure Studies on Dendrobium officinale[J]. Bulletin of Botanical Research, 2019, 39(4): 613-618.

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冰冻切片技术在铁皮石斛显微结构上的应用

Cryo-sectioning Technique for Microstructure Studies on Dendrobium officinale

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